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Updated: Jul 12, 2025
06:18
Imaging of Podocytic Proteins Nephrin, Actin, and Podocin with Expansion Microscopy
Published on: April 23, 2021
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Expansion Microscopy of Ciliary Proteins.
Sohyeon Park1, Xiaoyu Shi2,3,4
1Center for Complex Biological Systems, University of California Irvine, Irvine, CA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|October 19, 2023
Summary
Label-retention expansion microscopy (LR-ExM) enhances cellular imaging by expanding samples within a hydrogel. This method minimizes fluorescence loss, enabling high-resolution study of structures like primary cilia.
Area of Science:
- Cell biology
- Microscopy techniques
- Biochemistry
Background:
- Standard expansion microscopy (ExM) suffers from fluorescence loss and protein denaturation.
- These limitations hinder high-resolution imaging of cellular structures.
Purpose of the Study:
- To describe a detailed procedure for Label-retention expansion microscopy (LR-ExM).
- To demonstrate LR-ExM's utility in studying ciliary proteins at molecular resolution.
Main Methods:
- Embedding cells/tissues in a swellable hydrogel for sample expansion.
- Utilizing trifunctional anchors to preserve proteins during polymerization and denaturation.
- Introducing fluorophores post-expansion to minimize fluorescence loss.
Main Results:
- LR-ExM achieves high-resolution imaging with conventional fluorescence microscopes.
- The technique significantly improves signal-to-noise ratio compared to previous ExM methods.
- Molecular-scale structural details of primary cilia can be resolved.
Conclusions:
- LR-ExM is a robust method for high-resolution cellular imaging.
- It overcomes key limitations of standard expansion microscopy.
- LR-ExM enables detailed investigation of ciliary protein structures.
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